title: Single-molecule atomic force microscopy unravels the binding mechanism of a Burkholderia cenocepacia trimeric autotransporter adhesin.
abstract: Trimeric autotransporter adhesins (TAAs) are bacterial surface proteins that fulfil important functions in pathogenic Gram-negative bacteria. Prominent examples of TAAs are found in Burkholderia cepacia complex, a group of bacterial species causing severe infections in patients with cystic fibrosis. While there is strong evidence that Burkholderia cenocepacia TAAs mediate adhesion, aggregation and colonization of the respiratory epithelium, we still know very little about the molecular mechanisms behind these interactions. Here, we use single-molecule atomic force microscopy to unravel the binding mechanism of BCAM0224, a prototype TAA from B. cenocepacia K56-2. We show that the adhesin forms homophilic trans-interactions engaged in bacterial aggregation, and that it behaves as a spring capable to withstand high forces. We also find that BCAM0224 binds collagen, a major extracellular component of host epithelia. Both homophilic and heterophilic interactions display low binding affinity, which could be important for epithelium colonization. We then demonstrate that BCAM0224 recognizes receptors on living pneumocytes, and leads to the formation of membrane tethers that may play a role in promoting adhesion. Collectively, our results show that BCAM0224 is a multifunctional adhesin endowed with remarkable binding properties, which may represent a general mechanism among TAAs for strengthening bacterial adhesion.
authors.affiliation: Université catholique de Louvain, Institute of Life Sciences, Croix du Sud, 1, bte L7.04.01., B-1348, Louvain-la-Neuve, Belgium.
authors.forename: Sofiane
authors.initials: S
authors.lastname: El-Kirat-Chatel
authors.forename: Dalila
authors.initials: D
authors.lastname: Mil-Homens
authors.forename: Audrey
authors.initials: A
authors.lastname: Beaussart
authors.forename: Arsenio M
authors.initials: AM
authors.lastname: Fialho
authors.forename: Yves F
authors.initials: YF
authors.lastname: Dufrêne
mesh_terms: D018829:Adhesins, Bacterial / Q000378:metabolism* / Q000648:ultrastructure*; D056809:Alveolar Epithelial Cells / Q000382:microbiology; D001422:Bacterial Adhesion; D057508:Burkholderia cenocepacia / Q000502:physiology*; D002460:Cell Line; D003094:Collagen / Q000378:metabolism; D006801:Humans; D018625:Microscopy, Atomic Force; D011485:Protein Binding; D055503:Protein Multimerization
chemical_list: D018829:Adhesins, Bacterial; D003094:Collagen